Journal: Molecular Medicine Reports

Article Title: Overexpression of TIMP3 inhibits discogenic pain by suppressing angiogenesis and the expression of substance P in nucleus pulposus

doi: 10.3892/mmr.2020.10922

Figure Lengend Snippet: TIMP3, CD34 and substance P expression in rat NP tissue. Rats were percutaneously punctured with a 21G needle in coccygeal vertebra (puncture and TIMP3+puncture group). For TIMP3+puncture group, rats were injected with adenovirus vector (1×10 9 pfu/level) immediately after puncture. At day 28 after puncture, nucleus pulposus (NP) tissues were isolated for immunohistochemical staining. (A and B) Immunohistochemical staining was performed to measure TIMP3, CD34 and substance P expression in rat NP tissue. Overexpression of TIMP3 could reduce CD34 and substance P expression in NP tissue compared with the puncture group. Black arrow, positive staining. Data are presented as the mean ± SD. *P<0.05. Scale bar, 20 µm. TIMP3, tissue inhibitor of metalloproteinase-3.

Article Snippet: Primary antibodies against TIMP3 (ab155749, Abcam, Cambridge, UK), collagen-2 (ab34712, Abcam), GAPDH (ab181603, Abcam), Substance P (ab14184, Abcam), aggrecan (ab36861, Abcam) and CD34 (50589-R013, Sino Biological, Beijing, China) were used in the study.

Techniques: Expressing, Injection, Plasmid Preparation, Isolation, Immunohistochemical staining, Staining, Over Expression

Journal: Acta Pharmacologica Sinica

Article Title: Brain protection against ischemic stroke using choline as a new molecular bypass treatment

doi: 10.1038/aps.2015.104

Figure Lengend Snippet: Effects of oral choline treatment on angiogenesis in the ischemic brains. (A) Macroscopic images of the pial vasculature. (B) Microscopic images of ischemic cerebral cortex sections immunohistochemically stained using an anti-CD34 antibody to analyze vessel density, scale bar=20 μm. (C) Vessel density (n=6). (D) Number of microvessels in the cortex (n=3). Mean±SEM. bP<0.05, cP<0.01 vs sham; eP<0.05, fP<0.01 vs pMCAO.

Article Snippet: The capillary density was examined using a primary rabbit anti-rat CD34 antibody (Bioss, Beijing, China).

Techniques: Staining

Journal: Scientific Reports

Article Title: New lymphatic cell formation is associated with damaged brain tissue clearance after penetrating traumatic brain injury

doi: 10.1038/s41598-021-89616-3

Figure Lengend Snippet: Immunohistochemical staining images showing CD34 expression in ( A ) normal mouse brain tissue and ( B – F ) mouse brain tissue on days 3, 7, 15, 21 and 30 days, following the pTBI. ( A ) CD34 was not expressed in normal mouse brain tissue. CD34-positive cells and blood vessels (brown; black arrows) became visible at the injury site (red arrow) on days ( B ) 3 and ( C ) 7. ( D ) After 15 days, CD34 expression at the injury site (red arrow) decreased. ( E ) CD34 expression at the injury site could not be detected on day 21. Instead, black hemosiderin particles (black arrows) were observed at the injury site. ( F ) Some black hemosiderin particles remained visible at the injury site on day 30. ( G ) Immunohistochemical results of CD34 were analyzed by Image-Pro Plus 6.0 software. The average density (IOD) of CD34-positive cells in normal mouse brain tissue and in mouse brain tissue on days 3, 7, 15, 21 and 30 days, following pTBI. The n = 10/group was used for calculating the average IOD. Comparisons between groups were performed using the Kruskal–Wallis H test followed by Bonferroni post-hoc analysis.

Article Snippet: The sections were first incubated with rabbit anti-mouse PROX1 antibody (1:100; Boster Biological Technology Co., Wuhan, China), rabbit anti-mouse CD34 antibody (1:100; BosterBiological Technology) or goat anti-mouse LYVE-1 antibody (1:100; R&D Systems, Shanghai, China) at 4 °C for 24 h. Next, the sections were incubated with horseradish peroxidase-labeled goat anti-rabbit IgG or rabbit anti-goat IgG secondary antibodies.

Techniques: Immunohistochemical staining, Staining, Expressing, Software

Journal: Scientific Reports

Article Title: New lymphatic cell formation is associated with damaged brain tissue clearance after penetrating traumatic brain injury

doi: 10.1038/s41598-021-89616-3

Figure Lengend Snippet: Double immunofluorescence staining images showing CD34/LYVE-1 expression in the brain tissue 3 days after pTBI. Expression of ( A ) LYVE-1 (green); ( B ) CD34 (red); and ( C ) merge (yellow).

Article Snippet: The sections were first incubated with rabbit anti-mouse PROX1 antibody (1:100; Boster Biological Technology Co., Wuhan, China), rabbit anti-mouse CD34 antibody (1:100; BosterBiological Technology) or goat anti-mouse LYVE-1 antibody (1:100; R&D Systems, Shanghai, China) at 4 °C for 24 h. Next, the sections were incubated with horseradish peroxidase-labeled goat anti-rabbit IgG or rabbit anti-goat IgG secondary antibodies.

Techniques: Double Immunofluorescence Staining, Expressing

Journal: Frontiers in Neuroscience

Article Title: Trihydroxyethyl Rutin Provides Neuroprotection in Rats With Cervical Spinal Cord Hemi-Contusion

doi: 10.3389/fnins.2021.759325

Figure Lengend Snippet: CD34 immunohistochemical staining. (A) Sham group; (B) injury group; (C) T50 group; (D) T100 group; (E) statistical results of microvascular density showed that MVD of injury group, T50 group and T100 group were significantly lower than Sham group, and the MVD of T50 group and T100 group were significantly higher than injury group. * P < 0.05, comparison between each group and sham group; # P < 0.05, comparison between T50 group or T100 group and injury group.

Article Snippet: The resulting sections were soaked with 3% hydrogen peroxide to block endogenous peroxidase activity and then incubated overnight with rabbit polyclonal anti-CD34 antibody (1:1,000; Bioworld Technology, St. Louis Park, MN, United States).

Techniques: Immunohistochemical staining, Staining

Journal: Journal of Tissue Engineering and Regenerative Medicine

Article Title: In vivo analysis of vascularization and biocompatibility of electrospun polycaprolactone fibre mats in the rat femur chamber

doi: 10.1002/term.2868

Figure Lengend Snippet: HE staining, van Gieson staining, and immunohistochemical detection of CD 34: Representative histological stainings (a–o) 20 days after implantation into the femur chamber in Lewis rats. (a–c) HE staining of (1) granulation tissue, (2) implant, and (3) bone. (d–f) HE staining with higher magnification. (g–i) Collagen fibres were detected by van Gieson staining. (j–l) The presence of endothelial cells and accordingly the presence of vascular structures were confirmed by immunohistochemical detection of CD34. (m–o) Negative controls. Areas of the implant are marked (#), and arrows denote vascular structures [Colour figure can be viewed at wileyonlinelibrary.com ]

Article Snippet: For detection of capillaries, endothelial cells were immunohistochemically stained using a rabbit anti‐rat CD34 antibody (Hoelzel Diagnostika Handels GmbH, Koeln, Germany).

Techniques: Staining, Immunohistochemical staining